Influência da laserterapia na proliferação de células-tronco do ligamento periodontal humano

Low level laser irradiation (LLLI) has been used in Dentistry to promote wound healing and tissue regeneration. The literature shows a positive effect of LLLI on cell proliferation, but little is known about their effectiveness in promoting stem cells proliferation. The aim of this study was to evaluate the effect of LLLI on the proliferative rate of human periodontal ligament stem cells. Extracts of periodontal ligament were isolated from two third molars removed by surgical and/or orthodontic indication. After enzymatic digestion, the cells were grown in α-MEM culture medium supplemented with antibiotics and 15% fetal bovine serum. On the third subculture, the cells were irradiated with a InGaAlP-diode laser, using two different energy densities (0,5J/cm 2 - 16 seconds and 1,0J/cm² - 33 seconds), with wavelength of 660nm and output power of 30mW. A new irradiation, using the same parameters, was performed 48h after the first. A control group (non irradiated) was kept under the same experimental culture conditions. The Trypan blue exclusion test and the mitochondrial activity of the cells measured by MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] essay were performed to assess the cell proliferation in the intervals of 0, 24, 48 e 72 h after irradiation. The data of cell counts were submitted to nonparametrical statistical tests (Kruskal-Wallis and Mann-Whitney), considering a confidence interval of 95%. DAPI (4 -6-Diamidino-2-phenylindole) staining of the cells was performed at 72h interval to evaluate possible nuclear morphological changes induced by LLLI. The results of this study show that the energy density of 1,0 J/cm² promoted greater cell proliferation compared to the other groups (control and 0,5 J/cm²) at intervals of 48 and 72h. The mitochondrial activity measured by MTT essay showed similar results to the Trypan blue cell counting test. The group irradiated with 1,0J/cm² exhibited a significantly higher MTT activity in the intervals of 48 and 72h, when compared to the group irradiated with 0,5J/cm². No nuclear morphological change was observed in the cells from the three groups studied. It is concluded that LLLI has stimulatory effects on the proliferation of human periodontal ligament stem cells. Therefore, the use of laser irradiation in this cell type may be important to promote future advances in periodontal regeneration

Análise imunoistoquímica após terapia fotodinâmica com cloro-alumínio ftalocianina em tecidos gengivais de humanos

Although photodynamic therapy have been used as a useful tool over the past 30 years in oncology, few clinical trials have been conducted in dentistry. Photodynamic therapy (PDT) uses non - toxic photosensitizers and selective which are administered in target cells followed by local application of visible light, producing reactive oxygen species capable of causing cell death by apoptosis or necrosis, injured the local vasculature, and exert important effects on the im mune system. New generations of photosensitizing agents, such as nanoparticulate phthalocyanines, has shown excellent results in antitumor and antibacterial activity . In this context, the present work constitutes the first clinical protocol of local appli cation of nanoemulsion chloro - aluminum phthalocyanine (AlClFc) followed by irradiation in human gingiva, and analyzed descriptively and comparatively , by means of immunohistochemistry , the expression of RANK , RANKL , OPG and VEGF in a split - mouth model . Eight healthy volunteers with clinical indication for extraction were included in the study . Seven days before the extraction, was injected in the gingiva of participants, 5 μ M of nanoemulsion AlClFc followed by irra diation with diode laser (660nm , 7 J/cm2 ), the contralateral side was used as control. Tissue specimens were removed seven days after the TFD is performed. Tissues sample were divided into two groups (test and con trol groups) for histological and immunohistochemical analysis. Patients were monitored at days, 0, 7, 14 and 30 to assess adverse effects of the therapy. Vascular alterations were seen in gingival samples that received PDT. Areas of edema and vascular con gestion, and intense vascularization were viewed . Additionally, dystrophic calcification in subepithelial region were observed in the test group. The results showed a similar pattern of immunostaining scores of RANK, RANKL and VEGF between the test and co ntrol groups, with no statistically significant difference (p = 0.317, p = 0.777, p = 0 .814, respectively). RANK and RANKL exhibited weak or absent immunostaining in most specimens analyzed. There was n o immunostaining for OPG. VEGF showed moderate to stro ng immunostaining in specimens from the test group. In addition, the clinical study showed that therapy was well tolerated by all patients. Adverse effects were short - time and completely reversible. Taken together, the results presented in this study showe d that PDT mediated by nanoemulsion containing AlClPc is safe for clinical application in gingival tissue and suggests that a strong immunostaining for VEGF after therapy .

Efeitos da terapia fotodinâmica com aluminio – cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05). GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects.

Efeitos da laserterapia na atividade biológica de células de carcinoma epidermóide de língua humano

The low level laser therapy (LLLT) has shown to be effective in promoting the proliferation of different cells in vitro, including keratinocytes, osteoblasts, endothelial cells and stem cells. It has been speculated that the biostimulatory effect of LLLT could cause undesirable enhancement of tumor growth in neoplastic diseases, since the malignant cells are more susceptible to proliferative stimuli. Within this context, this study evaluated the effect of LLLT on epidermoid carcinoma of the tongue cell line (SCC25) proliferation and invasion. Cultured cells were irradiated with an InGaAIP diode laser, 660nm, 30mW using two energy densities (0.5J/cm2 and 1.0J/cm2). Proliferative activity was assessed through trypan blue staining method and through cell cycle analysis using flow cytometry. The invasive potential was measured through cell invasion assay using matrigel. Cyclin D1, E-cadherin, -catenin and MMP-9 expressions were analyzed by immunofluorescence and flow cytometry and related to the investigated biological activities. Proliferation curve demonstrated that SCC25 irradiated with 1.0J/cm2 had the highest proliferative rate when compared to the control group and the group irradiated with 0.5J/cm2 (p<0.05). LLLT affected cell cycle distribution and energy density of 1.0 J/cm2 promoted a higher percentage of cells in S/G2/M phases, with statistically significant differences at 24h interval (p<0.05). LLLT, mainly with 1.0J/cm2, revealed significantly higher potential for invasion and influenced the expression of cyclin D1, E-cadherin, -catenin and MMP-9, promoting the malignant phenotype. In conclusion, our results indicate that LLLT has an important stimulatory effect on proliferation and invasion of SCC25 cells, likely due to altered expression of proteins associated with these processes

Efeitos agudos da aplicação do laser de baixa potência sobre o desempenho neuromuscular do sóleo em sujeitos saudáveis

A laserterapia de baixa potência (LBP) tem demonstrado recentemente ser capaz de aumentar a resistência à fadiga, bem como potencializar o desempenho neuromuscular através de seus efeitos metabólicos e fotoquímicos. Estudos anteriores mostraram que o LBP reduziu o estresse oxidativo do exercício, promovendo um retardo da fadiga muscular e minimizando seus efeitos deletérios. O objetivo desse estudo foi avaliar se a aplicação do LBP antes de um protocolo de fadiga teria efeito sobre o desempenho neuromuscular nas variáveis eletromiográficas e dinamométricas do músculo sóleo em sujeitos saudáveis. Esta pesquisa caracterizou-se como um estudo experimental do tipo ensaio clínico controlado, randomizado e cego, no qual participaram 60 voluntários de ambos os sexos, com faixa etária entre 18 e 28 anos e fisicamente ativos, segundo o questionário internacional para a prática de atividade física (IPAQ). Os sujeitos foram alocados randomicamente em três grupos: Controle (G1), Placebo (G2) e Laser (G3) e todos os voluntários foram submetidos a uma avaliação inicial (AV-1), um Protocolo de Fadiga e uma avaliação final (AV-2), compostas por contrações isocinéticas para flexão plantar a uma velocidade de 90°/s. Os resultados encontrados nesse estudo mostraram que não houve diferença estatística entre os três grupos da pesquisa nas variáveis eletromiográficas de RMS e Frequência Mediana, contudo, em relação à dinamometria, o grupo que recebeu a aplicação do Laser obteve um índice de fadiga significativamente menor (p=0,04) quando comparado aos grupos Controle e Placebo. Além disso, a aplicação do laser pré-exercício também resultou em um aumento nas variáveis de desempenho potência e trabalho. Com isso, pode-se concluir que o LBP foi capaz de melhorar o desempenho do sóleo em sujeitos saudáveis, justificado pelo aumento da resistência à fadiga

Avaliação da efetividade de estratégias não farmacológicas para o alívio da dor de parturientes na fase ativa do período de dilatação no trabalho de parto

This study was aim to evaluate the effectiveness of strategies non pharmacological for the relief of the intensity of the parturient pain in the phase activates of the dilation period in the labor. Is a clinic rehearse of the type therapeutic intervention before and after" with a quantitative approach, accomplished in the Humanized Unit of Childbirth of the Maternity Januário Cicco School of the Federal University of Rio Grande do Norte, in Natal/RN, whit 130 parturient, being 30 in the pre-test of the strategies and 100 in the application of the strategies non pharmacological combined (breathing exercises, muscular relaxation and lombossacral massage) and isolated (shower bath in a normal temperature). We used the visual analogical scale to evaluate the intensity of the pain of the study parturient before and after" to the application of the strategies in the phases of acceleration, maximum inclination and desaceleration in the phase activates of dilation period in the labor. The principal results showed that the majority of the study parturient was between 20 to 30 years old (60%); with incomplete fundamental teaching (85%); family income until 2 minimum wages (74%); 78% had a companion and these, 44% were the own husband. The oxytocin was administered in the parturient during the phase activates of the labor in 81% of the cases and only 15% these women didn´t reciev anything medication. We verified significant relief (ρ=0.000) of intensity of the pain of the study parturient after application of the strategies non pharmacological combined and isolated. We concluded that the strategies non pharmacological combined and isolated were effective in the relief of the pain of the study parturient in the phase activates de labor

Compósito reforçado com sisal para aplicação na moda

The objective of this research is the fabrication of a composite reinforced with dyed sisal fiber and polyester matrix for application in the fields such as, fashion, clothing, interior textiles; fashion accessories are some of the examples. For the fabrication of the composite, the sisal fibers were subjected to processes such as: chemical treatment with sodium hydroxide (NaOH) in the removal of impurities; bleaching for removing the yellowish color of the natural fiber and dyeing with direct dyes to confer the colors blue, green and orange. The search for new technologies ecologically correct has become a major concern in recent decades. Studies show that composite polymer reinforced by natural fibers is suitable for a large number of applications, and its use is advantageous in terms of economic and ecological. The dyed fibers were cut to a length of 30 mm, is used in the confection of webs. For this purpose, a web preparer by immersion, developed in the Laboratory of Chemical Textile of UFRN. The composite sheets measuring 300 x 300 x3 mm were molded by compression, with unsaturated orthophthalic polyester as matrix, and the samples in sizes 150 x 25 x 3 mm were cut with the aid of a laser machine, to be subjected to traction and flexion. The mechanical properties of traction and flexion in three points were performed in the Laboratory of metal and mechanical tests of Materials Engineering of UFRN. The resulting samples from the tests were evaluated in scanning electron microscope (SEM) at CTGas RN. On the basis of the analysis of the results from the mechanical tests, it was observed that the composite had good mechanical behavior, both in traction as in flexion. Furthermore, it was observed that in the water absorption test, the samples had a different percentage among themselves, this occurred due to the variation of density found in the fibre webs. The images of the SEM showed the failures from the manufacturing process and the adhesion of fibre/matrix. When the samples were prepared with the dyed fibers to be applied in fashion, the results were positive, and it can be concluded that the main objective of this work was achieved

Avaliação clínica do efeito da irradiação pós-opratória do laser de baixa intensidade na cicatrização de gengivoplastias em humanos

Many studies have been made to understand the process of tissular cicatrization, as well as the possible effects of laser therapy in the wound healing. However, the influence of low frequency laser irradiation in the repairing process is not completely understood. Our study has the purpose to assess clinically the effect of postoperative irradiation of the low frequency laser in humans, and the gingival repairing process postgingivoplasty performed with the extern bevel technique. Twenty-four patients with inflammatory gingival hyperplasia were enrolled in this study, which did not reduce with basic periodontal procedures, and patients with melanin pigmentation, with esthetic indications. After surgery the test group, randomly selected by a drawing, received laser application with energy density of 4 J/cm2, immediately after surgery and each 48 hours, during a week, with a total of 4 sections. The control group did not receive irradiation. The visual clinical analyses were performed by a single blind examiner, in the 2nd, 4th, 6th, 8th, 15th and 21st days post surgery. For statistic analyses of the data was used a Q-square test. Concerning the color, the results showed a better wound healing during days 6 to 8. when assessed the degree of progress of surgical wound, the results showed that the test group had a better cicatrization compared with the control group in the 2nd, 6th, 8th and 15th days post surgery, and at the 21st day both groups had the same results. Our results confirm that the laser had clinical influence in the repairing process after gingivoplasty surgery during days 2 to 15 post surgery

Estudo da incorporação de resíduos de perfuração para aplicação em cimentação de poços de petróleo

During the drilling of oil and natural gas are generated solid waste, liquid and gaseous. These solid fragments, which are known as cuttings, are carried to the surface through the drilling fluid. Furthermore, this fluid serves to cool the bit, keeping the internal pressure of the well, and others. This solid residue is very polluting, because it has incorporated beyond the drilling fluid, which has several chemical additives harmful to the environment, some heavy metals that are harmful to the environment, such as lead. To minimize the residue generated, are currently being studied numerous techniques to mitigate the problems that such waste can cause to the environment, like addition of cuttings in the composition of soil cement brick masonry construction, addition of cuttings on the clay matrix for the manufacture of solid masonry bricks and ceramic blocks and coprocessing of the cuttings in cement. So, the main objective of this work is the incorporation of cuttings drilling of oil wells, the cement slurry used in the cementing operation of the well. This cuttings used in this study, arising from the formation Pendências, was milled and separated in a sieve of 100 mesh. After grinding had a mean particle sike in order of 86 mm and crystal structure containing phases of quartz and calcite type, characteristic of the Portland cement. Were formulated and prepared slurries of cement with density 13 lb / gal, containing different concentrations of gravel, and realized characterization tests API SPEC 10A and RP 10B. Free water tests showed values lower than 5.9% and the rheological model that best described the behavior of the mixtures was the power. The results of compressive strength (10.3 MPa) and stability (Dr <0.5 lb / gal) had values within the set of operational procedures. Thus, the gravel from the drilling operation, may be used as binders in addition to Portland cement oil wells, in order to reuse this waste and reduce the cost of the cement paste.